Primary Cell KO Pools: Advance your immunology research with edited-to-order primary cells

Get Guaranteed Editing Results in Primary Cells

Primary Cell KO Pools are made-to-order knockouts with a guaranteed editing efficiency of >80% in human primary cells generated in 2 weeks or faster. EditCo consistently achieves >90% editing efficiency, creating cell pools that are functionally similar to a clone. As primary cell clones are biologically difficult to produce, EditCo’s high editing efficiency provides a unique approach to generating cell populations of interest.

Whether you are leveraging CRISPR for validating targets for drug discovery or modeling a biological process for the investigation of a novel gene of interest, failed experiments lead to a loss of time and resources. EditCo’s Primary Cell KO Pools are the solution for obtaining quick and reliable functional data while reducing the risk for false negatives in your critical assays.

Figure 1. Editing Efficiency and Viability for CD4+ T cell pools. Values represent the average of three different donors showing high editing efficiency, as determined by ICE analysis, across a variety of loci (blue bars, mean KO scores +- SEM) without affecting viability at the time of freezing (cyan dots).

Figure 2. Editing Efficiency leads to protein depletion as shown by flow cytometry (BD Symphony A1).

High viability and expansion of edited pools

The editing process and further expansion before freezing do not affect cell fitness. Edited cells can be thawed and expanded for any desired downstream assay, maintaining high cell viability and editing levels (Figure 3).

Figure 3. CD4+ pool stability after thaw. Panel A. PD1 expression in TCR activator-treated cultures peaked on day 3 in all tested donors (edited and mock samples). Isotype control staining over the same period is shown for reference. Panel B. Relative fold expansion for edited pools cultured with TCR activator or IL2 alone. Panel C. Pools of 4 different edits from 3 separate donors showed >90% viability and no decrease in editing efficiency after 14 days in culture.

High functionality

Edited CD4+ T-cells produced levels of IFNg, IL2 and TNFa after mitogenic stimulation that correlated with known target gene function.

Figure 4. Cytokine secretion in edited cell pools following cell expansion. Edited cells from 3 donors   were thawed and then stimulated with PMA/ionomycin for 6 hours. Supernatants were harvested and measured for concentrations of IFNG (left panel), IL2 (center panel) and TNFA (right panel) as well as IL10 and IL5 (not shown) by FACS-ELISA (Miltenyi).  Values were quantile normalized to reduce batch effects. Following ANOVA, Tukey’s multiple comparison test was performed between Mock and each gene target to determine statistical significance. (*) p<0.05; (**) p<0.01; (***) p<0.005; (****) p< 0.0001.

Figure 5. Intracellular Cytokine measurement in edited cell pools. Knockout CD4+ T cell pools of IFNg, IL2 or TNFa were thawed and stimulated with PMA/ionomycin for 6 hours. Cells were stained for CD3, CD4 and viability, permeabilized and then stained for presence of IFNG, IL2 or TNFA. Production of either cytokine in CD3/CD4/Live gated events for each Gene Target knockout culture (Bottom histograms) was compared to unedited control (Mock, Middle histograms) as well as TRAC knockout (Top histograms).